Inhibition of Metastatic Lung Cancer in C57BL/6 Mice by Marine Mangrove Rhizophora apiculata

  • Prabhu, V. Vinod (Department of Biotechnology, Karunya University) ;
  • Guruvayoorappan, C. (Department of Biotechnology, Karunya University)
  • Published : 2013.03.30


Metastasis is one of the hallmarks of malignant neoplasms and is the leading cause of death in many cancer patients. A major challenge in cancer treatment is to find better ways to specifically target tumor metastasis. In this study, the anti-metastatic potential of the methanolic extract of Rhizophora apiculata (R.apiculata) was evaluated using the B16F-10 melanoma induced lung metastasis model in C57BL/6 mice. Metastasis was induced in C57BL/6 mice by injecting highly metastatic B16F-10 melanoma cells through the lateral tail vein. Simultaneous treatment with R.apiculata extract (10 mg/kg b.wt (intraperitoneal) significantly (p<0.01) inhibited pulmonary tumor nodule formation (41.1 %) and also increased the life span (survival rate) 107.3 % of metastatic tumor bearing animals. The administration of R.apiculata extract significantly (p<0.01) reduced biochemical parameters such as lung collagen hydroxyproline, hexosamine, uronic acid content, serum nitric oxide (NO), ${\gamma}$-glutamyl transpeptidase (GGT) and sialic acid levels when compared to metastasis controls. These results correlated with lung histopathology analysis of R.apiculata extract treated mice showing reduction in lung metastasis and tumor masses. Taken together, our findings support that R.apiculata extract could be used as a potential anti-metastasis agent against lung cancer.


  1. Aragon-Ching JB, Zujewski J (2007). CNS metastasis: an old problem in a new guise. Clinical Cancer Res, 13, 1644-7.
  2. Ambs S, Merriam WG, Bennett WP, et al (1998). Frequent nitric oxide synthase-2 expression in human colon adenomas: implication for tumor angiogenesis and colon cancer progressionm. Cancer Res, 58, 334-41.
  3. Antony JJ, Sivalingam P, Siva D, et al (2011). Comparative evaluation of antibacterial activity of silver nanoparticles synthesized using Rhizophora apiculata and glucose. Colloids Surf B Biointerfaces, 88, 134-40.
  4. Bergman I, Loxley R (1970). The determination of hydroxyproline in urine hydrosylates. Clin. Chem Acta, 27, 347-9.
  5. Bhatia S, Tykodll SS, Thompson JA (2009). Treatment of metastatic melanoma: an overview. Oncology (Williston Park), 23, 488-96.
  6. Bhavanandhan VP, Katlic AW, Banks J, et al (1981). Partial characterization of sialoglycopeptides produced by cultured human melanoma cells and melanocytes. Biochemistry, 20, 3586-94.
  7. Bitter T, Muir HM (1962). A modified uronic acid carbazole reaction. Anal. Biochem, 4, 330-34.
  8. Branco-Price C, Zhang N, Schnelle M, et al (2012). Endothelial cell HIF-$1{\alpha}$ and HIF-$2{\alpha}$ differentially regulate metastatic success. Cancer Cell, 21, 52-65.
  9. Carretero J, Obrador E, Anasagasti MJ, et al (1999). Growthassociated changes in glutathione content correlate with liver metastatic activity of B16 melanoma cells. Clin Exp Metastasis, 17, 567-74.
  10. Chambers AF, Groom AC, MacDonald IC (2002). Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer, 2, 563-72.
  11. Cole SP (1986). Rapid chemosensitivity testing of human lung tumor cells using the MTT assay. Cancer Chemother Pharmacol, 17, 259-63.
  12. Edwards P, Cendan JC, Topping DB, et al (1933). A colourimetric method for determination of glucosamine. J Biochem, 27, 1824-8.
  13. Edwards P, Cendan JC, Topping DB, et al (1996). Tumor cell nitric oxide inhibits cell growth in vitro, but stimulates tumorigenesis and experimental lung metastasis in vivo. J Surg Res, 63, 49-52.
  14. Engbrin JA, Hoffman MP, Karmand AJ, Kleinman HK (2002). The B16F-10 receptor for a metastasis promoting site on Laminin-1 is a heparan sulfate/ chondroitin sulfate containing proteoglycan. Cancer Res, 62, 3549-54.
  15. Fukumura D, Kashiwagi S, Jain RK (2006). The role of nitric oxide in tumor progression. Nat Rev Cancer, 6, 521-34.
  16. Fuster MM, Esko JD (2005). The sweet and sour of cancer:Glycans as novel therapeutic targets. Nature Reviews Cancer, 5, 526-42.
  17. Green LC, Wagner DA, Glogowski J, et al (1982). Analysis of nitrate, nitrite and (15N) nitrate in biological fluids. Anal Biochem, 126, 131-8.
  18. Gupta GP, Massague J (2002). Cancer metastasis: building a framework. Cell, 127, 679-95.
  19. Herzig M, Christofori G (2002). Recent advances in cancer research: mouse models of tumorigenesis. Biochim Biophys Acta, 1602, 97-113.
  20. Hunter KW, Crawford NPS, Alsarraj J (2008). Mechanisms of metastasis. Breast Cancer Res, 10, 2.
  21. Hyoudou K, Nishikawa M, Umeyama Y, et al (2004). Inhibition of metastatic tumor growth in mouse lung by repeated administration of polyethylene glycol-conjugated catalase: quantitative analysis with firefly luciferase-expressing melanoma cells. Clin Cancer Res, 15, 7685-91.
  22. Khan N, Mukhtar H (2010). Cancer and metastasis: prevention and treatment by green tea. Cancer Metastasis Rev, 29, 435-45.
  23. Klein CA (2008). Cancer. The metastasis cascade. Science 321,1785-87.
  24. Kuttan R, Bhanumathy P, Nirmala K, George MC (1985). Possible anticancer activity of turmeric. Nutr Cancer, 29, 197-202.
  25. Lee JM, Yanagawa J, Peebles KA, et al (2008). Inflammation in lung carcinogenesis: new targets for lung cancer chemoprevention and treatment. Crit Rev Oncol Hematol, 66, 208-17.
  26. Leyon PV, Kuttan G (2004). Effect of Withania somnifera on B16F-10 meanoma induced metastasis in mice. Phytother Res, 18, 118-22.
  27. Leyon PV, Lini CC, Kuttan G (2005). Inhibitory effect of Boerhaavia diffusa on experimental metastasis by B16F10 melanoma in C57BL/6 mice. Lifesciences, 76, 133-49.
  28. Lipponen P, Aaltomaa S, Tammi R, Argen U, Kosma VM (2001). High stromal hyaluronan level is associated with poor differentiation and metastases in prostate cancer. Eur J Cancer, 37, 849-56.
  29. Luzzi KJ, MacDonald IC, Schmidt EE, et al (1998). Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early micrometastases. Am J Pathol, 153, 865-73.
  30. Mokkhasmit M, Swatdimongkol. K, Satrawaha P (1971). Study on toxicity of Thai medicinal plants. Bull Dept Med Sci. 12, 36-65.
  31. Nathan C (1992). Nitric oxide as a secretory product of mammalian cells. FASEB J, 6, 3051-64.
  32. Nelson DL, Cox MM (2005). Lehninger's Principles of Biochemistry, 4th Edition, W. H. Freeman and Company, New York.
  33. Nguyen DX, Massagué J (2007). Genetic determinants of cancer metastasis. Nat Rev Genet, 8, 341-52.
  34. Nonaka Y, Iwagaki H, Kimura T, Fuchimoto S, Orita K (1993). Effect of reactive oxygen intermediates on the in vitro invasive capacity of tumor cells and liver metastasis in mice. Int J Cancer, 54, 983-86.
  35. Prabhu VV, Guruvayoorappan C (2010). Nitric oxide: Pros and cons in tumor progression. Immunopharmacol Immunotoxicol, 32, 387-92.
  36. Prabhu VV, Guruvayoorappan C (2012). Evaluation of immunostimulant activity and chemoprotective effect of mangrove Rhizophora apiculata against cyclophosphamide induced toxicity in BALB/c mice. Immunopharmacol Immunotoxicol, 34, 607-14.
  37. Prabhu VV, Guruvayoorappan C (2012a). Anti-inflammatory and anti-tumor activity of the marine mangrove Rhizophora apiculata. J Immunotoxicology, 9, 341-352.
  38. Prabhu VV, Siddikuzzaman, Grace VM, Guruvayoorappan C (2012b). Targeting tumor metastasis by regulating Nm23 gene expression. Asian Pac J Cancer Prev, 13, 3539-48.
  39. Pradeep K (2002). Effect of piperine on the inhibition of lung metastasis induced B16F-10 melanoma cells in mice. Clinical & Experimental Metastasis, 19, 703-08.
  40. Premanathan M, Arakaki R, Izumi H, et al (1999). Antiviral properties of a mangrove plant, Rhizophora apiculata Blume, against human immunodeficiency virus. Antiviral Res, 44, 113-22.
  41. Schantz SP, Brown BW, Lira E, Taylor SL, Beddingfield N (1987). Evidence for the role of natural immunity in the control of metastatic spread of head and neck cancer. Cancer Immunology Immunotherapy, 25, 141-48.
  42. Schauer R (1985). Sialic acids and their role as biological masks. Trends Biochem Sci, 10, 357-60.
  43. Shi Q, Huang S, Jiang W, et al (1999). Direct correlation between nitric oxide synthase II inducibility and metastatic ability of UV-2237 murine fibrosarcoma cells carrying mutant p53. Cancer Res, 59 , 2072-75.
  44. Sithranga BN, Kandasamy K, Subramanian M, You-Jin J (2011). Effect of mangrove tea extract from ceriops decandra (Griff.) Ding Hou. on salivary bacterial flora of DMBA Induced hamster buccal pouch carcinoma. Indian J Microbiol, 51, 338-44.
  45. Skoza L, Mohos S (1976). Stable thiobarbituric acid chromophore with dimethyl sulphoxide. Application to sialic acid assay in analytical de o acetylation. Biochem J, 159, 457-62.
  46. Talmadge JE, Fidler IJ (2010). AACR centennial series: the biology of cancer metastasis: historical perspective. Cancer Res, 70, 5649-69.
  47. Tate SS, Meister A (1974). Inhibition of gamma glutamyl transpeptidase amino acids, peptides and derivatives and analogs of glutathione. J Biochem, 23, 1602.
  48. Thejass P, Kuttan, Girija (2006). Antimetastatic activity sulforaphane. Life Sci. 78, 3043-50.
  49. Vedolva E, Borovansky J (1994). Evaluation of serum sialic acid fractions as marker from malignant melanoma. Cancer Lett, 78, 171-5.
  50. Voet D, Voet JG (1995). Biochemistry, New York: John Wiley, 157-258.
  51. Wang YP, Wang MZ, Luo YR, Shen Y, Wei ZX (2012). Lentivirus-mediated shRNA interference targeting SLUG inhibits lung cancer growth and metastasis. Asian Pac J Cancer Prev, 13, 4947-51.
  52. West DC, Hampson IN, Arnold F, et al (1985). Angiogenesis induced by degradation products of hyaluronic acid. Science, 228, 1324-26.
  53. WHO glonocon report 2008.
  54. WHO Report (2008). National Policy on Traditional Medicine and Regulation of Herbal Medicines - Report of a WHO global survey.

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